WO2014185761A1

WO2014185761A1 - Novel 3,4-diaminopyridine composition for treating muscle fatigability associated with neurovegetative disorders

Info

Publication number: WO2014185761A1
Application number: PCT/MA2013/000041
Authority: WO
Inventors: Benziane HAMID
Original assignee: Université Mohammed V Souissi
Priority date: 2013-05-16
Filing date: 2013-12-05
Publication date: 2014-11-20
Also published as: MA35698B1

Abstract

The subject matter of the invention relates to a novel composition of 3,4-diaminopyridine (3,4-DAP) rendering same stable and homogeneous, prepared in the form of a capsule for the treatment and control of paraneoplastic Lambert-Eaton myasthenic syndrome, characterised by muscle fatigability associated with neurovegetative disorders. Said drug is used in fatigability associated with multiple sclerosis, botulism and congenital myasthenic syndromes and Alzheimer's disease. The novelty also relates to the dosing protocol of the active agent and of the carriers. In the novel 3,4-DAP composition, the mixture is supplied in the form of a capsule, tablet or oral suspension, or other dosage forms.

Description

Title: Novel composition of 3,4-diaminopyridine for the treatment of muscular fatigability associated with neurovegetative disorders

The description

The invention relates to a new composition of 3,4-diaminopyridine (3,4-dap) making it homogeneous stable and at 5 years, prepared in capsule form for treating and controlling the progression of the Lambert-Eaton myasthenic paraneoplastic syndrome, characterized by muscle fatigability associated with neurovegetative disorders.

This drug is also used in the fatigability of multiple sclerosis, botulism and congenital myasthenic syndromes and in Alzheimer's disorders. The medicinal treatment of pathologies of Lambert-Eaton myasthenic paraneoplastic syndrome, multiple sclerosis, botulism, congenital myasthenic syndromes and in Alzheimer's disorders, for which a beneficial effect has been reported, make use of the association plasmapheresis immunoglobulins, corticosteroids and azathioprine, This treatment is however restrictive and often poorly tolerated by patients; moreover the positive effect of plasmapheresis is unfortunately limited in time.

In addition, the use of the 3,4-diaminopyridine (3,4-dap) molecule often prescribed in these cases does not rely on a dosage defined in Lambert-Eaton syndrome. Bevert and Jost recommend the use of an average dosage of 40mg / d (15-60mg / d), in three to four doses.

Our study shows the interest of making magistral preparations adapted to the needs of each patient. It therefore appears necessary to make fine dose adjustments and to undertake larger studies to determine the reasons for this particular individual response. Our invention recommends the use of a compound characterized by a concentration of the active ingredient per dose in the range of 2.5 to 5 mg per dose four to eight times per day according to the fatigue felt by the patient.

3,4-dap is prescribed in second intention at a maximal dose which varies between 60 and 100mg / day, because of its numerous central undesirable effects (nauseas, anxiety, convulsions ...), peripheral (paresthesia essentially) and the autonomic nervous system (coldness of the extremities, palpitation, visual disturbances, cough, bronchial hypersecretion.Thus, this pharmaceutical compound may comprise other active principles.

Different excipients were tested (carboxy methyl cellulose, wheat starch, magnesium stearate and E-120 cochineal carmine), distinguishing two types of preparation: binary (2 excipients in addition to the active ingredient) and ternary (3 excipients in addition to the active ingredient). The capsules used are small (No. 4 corresponding to a volume of 0.22 ml). Since the 3,4-APD is made of a pyridine ring and two groups amin _e s, the dosage of the active ingredient was carried out on 10 capsules each batch by uv-visible spectrophotometry, ROUESSAC F., ROUESSAC A. 2000. The batch is considered compliant when the average content of active ingredient is between 85 and 115%. If more than 3 capsules are outside the 75-125% range, the lot is rejected (Deutscher Arzneimittei Codex 8th 1986. 3,4-Diaminopyridine, OD25).

The new composition of 3,4-DAP for the preparation of a pharmaceutical compound comprises:

carmine,

- wheat starch,

- carboxymethyl cellulose or magnesium stearate

specifying the grammage of each compound ensures the stability of the preparation over a period of 5 years at room temperature,

This new compound of formula 3,4-DAP and a pharmaceutically compatible salt thereof, as active ingredient for the preparation of a pharmaceutical composition for the treatment of muscle fatigability associated with neurovegetative disorders in the man.

The stability study of this preparation was carried out within the temperature range between 5 and 60 ° C, thus taking into account the usual conditions of conservation and those of accelerated aging [14]. For this we used the cold room of the pharmacy (5 ± 1 ° C), an oven with air flow (20-220 ° C ± 2 ° C). The stability tests carried out in our series did not reveal any degradation (SANDERS D.B., MASSEY J.M., SANDERS I.L., EDWARDS L.J.2000), significant of the finished product between 4 and 40 ° C over a period of 6 months.

These results do not seem to agree with those of Sanders, who in a clinical trial chose lactose as an excipient. The 3,4-diaminopyridine (3,4-dap + lactose) capsules at 10 and 20 mg were stored at -20 ° C; the choice of this method of preservation seems to avoid the active ingredient-excipient interaction. These storage difficulties can be avoided by replacing lactose with wheat starch and adding magnesium stearate to improve the rheological properties of the final mixture.

On his briefing note [AGEPS. Information note concerning the use of October 2002], the General Agency of Equipment and Health Products (AGEPS), without proposing a formula of preparation, draws the attention of the health professionals on the risk of interaction of the 3,4- dap with magnesium by a transmembrane ionic mechanism. The content of magnesium stearate per capsule is 1.5 mg on average. This quantity is very small to have a noticeable effect.

This new composition is not marketed industrially. We prepared it at the hospital and thus submitted to the status of hospital preparation. In a formulation test, the gelatin capsule form was chosen because of the characteristics of the active ingredient (crystalline powder), the simplicity of the manufacture and the ease of carrying out a dosage titration.

Preparation of 3,4-diaminopsrydine capsules

The choice of excipients was oriented towards raw materials whose particle size is close to that of 3,4-DAP. These excipients are supplied by a pharmaceutical establishment (lactose, carboxymethyl cellulose, wheat starch). The E-120 scale carmine was incorporated as a visual indicator of homogeneity of the powder. Magnesium stearate was then used to improve the flow. The capsules are envelopes in gelatin No. 4. The active ingredient was provided by the General Agency of Equipment and Health Products, excipients and capsules by the Cooper. As part of the Good Hospital Pharmacy Practices, the excipients were identified according to the methods described by the European Pharmacopoeia 4th edition. 3,4-DAP was identified according to the reactions described in the German Pharmacopoeia 8th edition (monograph 3,4-diaminopyridine).

The formulation tests concerned 0.22 ml qs capsules, prepared successively, from six different formulations:

• 3,4-DAP, carmine and lactose (Fl).

• 3,4-DAP, carmine and carboxy methyl cellulose (F2).

• 3,4-DAP, carmine and wheat starch (F3).

• 3,4-DAP, carmine and lactose + magnesium stearate (F4).

• 3,4-DAP, carmine and carboxymethyl cellulose + magnesium stearate (F5).

• 3,4-DAP, carmine and wheat starch + magnesium stearate (F6).

All these formulas are not effective, only the F6 shows a positive activity on the clinical signs of the diseases characterized by the fatigability.

The preparation of the capsules is based on a semi-automatic gelatin capsule that makes it possible to make a batch of 100 capsules at a time, according to the same procedure, with the same material. and by the same people. The dosage should be adapted to each patient, in the absence of the possibility of carrying out a therapeutic follow-up (tissue dosage of the drug), the dosage titration is done on the basis of the clinic and the electromyographic examination. The first administrations will use low-dose capsules (2.5 to 5 mg of active ingredient per capsule). These doses may be increased depending on the patient's tolerance [BEVER CT, ANDERSON PA, LESLIE J., PAMTCH HS, DHTB I, KHAN OA and] 1996 9 TIME RW, MASSEY JM, SANDERS DB 2000).

The new composition of 3,4-DAP varies between: 0.1 to 0.5 mg for carmine, 5 to 7 mg for wheat starch, and from there 2 mg for magnesium stearate for a capsule of 0.22 ml of volume ,

The new composition of 3,4-DAP must respect the succession of steps, first the dose of the active ingredient followed by the color indicator (Carmine COCHNILLE) then wheat starch and finally magnesium stearate. The homogenization mixing process respects the duration of 20 to 30 minutes.

Mass uniformity check

The mass uniformity control of the hard shell capsules was carried out by weighing 20 capsules individually using a Sartorius electronic scale (e = 1 mg, d = 0.1 mg) and software. parameterized according to the recommendations of the European Pharmacopoeia 4th edition (Capstool version 1.1, March 1998). It admits capsules whose average mass is between 40 mg and 300 mg, 10% of the limit difference of the average mass. The mass of any unit may not deviate by more than twice that percentage.

Control of the active ingredient content

Consisting of a pyridine ring and two amino groups (Figure 1), 3,4 DAP is in the form of a powder very sparingly soluble in water and ethanol, easily soluble in methanol and practically insoluble in the ether. Its structure gives it maximum absorption at 292 nm in methanolic medium (Figure 2).

Control of the content of this active ingredient is carried out on ten capsules, dosed individually, using a UV-Visible spectrophotometer (Cary 1 Bio Variant®). The contents of each capsule are dissolved in methanol for analysis (Carlo Erba®). The solution is filtered on a No. 4 paper filter (Wathman type), the filtrate is collected (solution S0). One ml of the solution S0 is then diluted in 10 ml of methanol (solution SI). The spectrum is recorded between 200 nm and 400 nm, with a scanning speed of 100 nm / min and a sampling interval of 2 nra. The absorbance reading is carried out at 292 nm _. A calibration line is established from three concentrations of 3,4-DAP at 40 mg / 1.20 mg / l and 10 mg / l. Repeatability and reproducibility were tested on ten samples. The excipient-active ingredient interference was analyzed by studying the absorbance spectra of different solutions of carmine and excipients.

The batch is considered compliant, when the average content of active ingredient is between 85-115%. When more than three capsules are outside this range and / or when more than one capsule is outside the 75% -125% range, the lot is rejected [European Pharmacopoeia 4th edition, Strasbourg 2002: 215.] .

This new protocol for the determination of the active ingredient uses UV-Vis spectrophotometry in methanolic medium, after filtration where the scanning speed is kept fixed. Mass uniformity control

The mass uniformity tests carried out on the six different formulations automatically removed mixtures containing only one excipient (in addition to carmine). These are the binary formulations (F1-F3):

• Fl: average mass of the content; 0.0877 g (min: 0.070 g, max: 0.1122 g). Three capsules are outside the ± 10% range (0.10 g, 0.1122 g, 0.070 g);

• F2: average mass of the contents: 1,256 g (min: 1,1262 g, max: 1,6012 g). Two capsules deviate by ± 20% (1.5080 g, 1.6012 g);

Control of the active ingredient content

The control of the content of the active ingredient is only envisaged for the formulations which comply with the plan of uniformity of mass: - Formulation F4: The test of the 3,4-DAP content in batch F4 shows an average content 3.05 mg (2.40-3.52 mg). Five capsules are outside the 85-115% range (2.12-2.87 mg) and three capsules are outside the 75-125% range.

Formulation F6: The individual dosage of ten capsules of 3,4-DAP taken at random on the semi-automatic capsule, shows an average content of 2.33 mg (1.90-2.33 mg). Three units are outside the 85-115% range [18] (1.90, 1.97 and 1.97 mg). No capsule is outside the 75-125% range [18].

Our compound characterized by a pharmaceutical composition which may further comprise one or more compatible toxicologically harmless excipients with a dosage of 2.5 mg Stability test

At temperatures between 4 ° C and 40 ° C

The stability study of the capsules of 3,4-DAP at 2,5 mg, between 4 ° C and 40 ° C, shows that their composition is not modified three months after their manufacture. Indeed :

The visual observation of the solutions prepared from said capsules, thus preserved for 3 months, did not reveal any abnormal coloration, precipitation or crystallization;

The absorption spectrum remains unchanged;

The UV-visible spectrophotometric assay of the active ingredient does not vary significantly (5%).

So we chose for our hospital preparation, a shelf life of two months between 4 ° C and 40 ° C.

At temperatures above 40 ° C, the 3,4-DAP capsules at 2.5 mg did not undergo significant degradation after two months of storage in different experimental temperatures up to 60 ° C. If the residence at 60 ° C is equal to or exceeds three months the powder contained in the capsules becomes brownish. The absorption spectrum is significantly modified and the average content per capsule of the principle decreases to 33% (1.67 mg).

Study of the stability of the finished product

We have not found in the literature data on the stability of 3,4-DAP, except for certain indications appearing on the actual packaging of this active ingredient (in particular a period of validity of two years, packaging in good faith). of brown glass, a List I label, with the words: "Keep the packaging carefully closed and protected from light"). Three lots of 100 capsules were used in this study. We therefore opted for a stability test within the temperature range of 5 to 60 ° C, taking into account the usual conditions of conservation and accelerated aging.

The stability test showed good preservation of the finished product within the extreme limits of the normal conditions of use. The clinical and electromyographic evaluation revealed a beneficial effect of this hospital preparation.

The contribution of our invention is that the stabilization process results from the 3,4 diaminopyridine formulation, E-120 cochineal carmine, wheat starch, magnesium stearate and lasts 5 years at room temperature (25 ° C). The new composition of 3,4-DAP the mixture is delivered in the form of capsule, tablet or oral suspension, or other galenic forms

Claims

The revendications

1. The novel composition of 3,4-DAP for the preparation of a pharmaceutical compound for the treatment of muscle fatigability associated with neurovegetative disorders in humans comprising:

carmine,

- wheat starch,

carboxymethyl cellulose or magnesium stearate

Characterized in that the new dosage of each compound ensures the stability of the preparation over a period of 5 years at room temperature,

2. The novel composition of 3,4-DAP according to claim 1, characterized in that the amount of the excipients varies between: 0.1 to 0.5 mg for carmine, 5 to 7 mg for wheat starch, and from there 2mg for magnesium stearate for one capsule of 0.22ml volume,

3. The new composition of 3,4-DAP according to claim 1 and 2, characterized in that the procedure respects the succession of steps, first the dose of the active ingredient followed by the color indicator (Carmine COCHNTLLE) then wheat starch and finally magnesium stearate,

4. The novel 3,4-DAP composition according to claim 1, characterized in that the homogenization mixing process has a duration of 20 to 30 minutes,

5. The novel composition of 3,4-DAP according to claim 1,2,3 and 4, characterized in that this mixture is delivered in capsule, tablet or oral suspension form.